用SDS-PAGE和Vorum银染法比较真菌菌丝体破壁效果  

Comparison of the Effect for Filamentous Fungal Breaking-Wall by SDS-PAGE and Vorum Silver Staining

作　　者：李敏惠[1] 潘克俭[2] 宋丽萍[2] 钟文英[2] 邹珂珂[2] 杨平[3] 

机构地区：[1]成都医学院科研实验中心,四川成都610083 [2]成都医学院生物医学系,四川成都610083 [3]成都医学院生物化学教研室,四川成都610083

出　　处：《生物技术通讯》2011年第6期867-869,共3页Letters in Biotechnology

基　　金：四川省教育厅青年基金(10ZB095);成都医学院创新性实验项目(CX2010015)

摘　　要：目的:研究不同处理方法对丝状真菌Pythium sp.GY1938菌丝体破壁效果的差异。方法:采用7种方法对真菌菌丝体进行破壁处理,将制备的胞内可溶蛋白质样品通过SDS-PAGE和Vorum法银染处理,比较不同破壁方法处理后蛋白质释放效果的优劣。结果:方法VI(石英砂、液氮、研磨)的破壁效果最好,蛋白质的分离效率和提取质量最高。结论:方法VI适合于丝状真菌Pythium sp.GY1938蛋白组学研究中目的蛋白的提取。Objective： To compare the effects in the release of the intracellular protein in Pythium sp.GY1938 strain.Methods： Seven kinds of treatment including grinding with silica sand,sonication after grinding with silica sand,sonication with silica sand,sonication,grinding with liquid nitrogen,grinding with silica sand and liquid nitrogen,sonication after grinding with silica sand and liquid nitrogen were used in this research.Then,each lysate was treated with SDS-PAGE and the efficiency was evaluated by Vorum silver staining.Results： The silver staining results showed that the sixth method,grinding with silica sand and liquid nitrogen,was the optimum mean to break cell wall of this fungi because of the high integrality and separation efficiency of protein extraction.Conclusion： The method grinding with silica sand and liquid nitrogen was the first selective intracellular protein extraction means in Pythium sp.GY1938 strain for the further fungal proteomic research.

关 键 词：丝状真菌 PYTHIUM sp.GY1938 破壁 银染 

分 类 号：Q949.32[生物学—植物学] Q51